Nerve-growth-factor-dependent and cell-density-independent survival of septal cholinergic neurons in culture from postnatal rats

• Published in: Neuroscience research 1990-06, Vol.8 (2), p.69-82
• Main Authors: Hatanaka, Hiroshi, Nishio, Chika, Kushima, Yoichi, Tsukui, Hiroko
• Format: Article
• Language: English
• Subjects: Acetylcholinesterase; Acetylcholinesterase - metabolism; Animals; Animals, Newborn - physiology; Biochemistry and metabolism; Biological and medical sciences; Brain - cytology; Cell Count; Cells, Cultured; Central nervous system; Choline acetyltransferase; Choline O-Acetyltransferase - metabolism; Female; Fundamental and applied biological sciences. Psychology; Immunohistochemistry; Male; Nerve Growth Factors - physiology; Neuronal survival; Neurons - physiology; Papain; Postnatal CNS neuron; Pregnancy; Rats; Rats, Inbred Strains; Sympathetic Nervous System - cytology; Vertebrates: nervous system and sense organs
• ISSN: 0168-0102; 1872-8111
• DOI: 10.1016/0168-0102(90)90060-R

We have established a primary neuronal cell culture technique from the postnatal (P11 to P15) rat CNS to study the nerve growth factor (NGF) response to basal forebrain cholinergic neurons. The survival of septal cholinergic neurons in culture was monitored both by the determination of choline acetyltransferase activity and by counting acetylcholinesterase-positive cells. Cells obtained from postnatal septal regions were found to require a plentiful oxygen supply during the dissociation of the cells. NGF-mediated survival of the septal cholinergic neurons was similarly observed in the cultures by using different plating cell densities up to 12.5 × 10 5 cells/cm 2. These results suggest that the promotion by NGF of cell survival in culture is independent of plating cell density.